This renewal application, in response to the RFA CA-04-006 "The Early Detection Research Network: Biomarker Developmental Laboratories" (EDRN/BDL), proposes an integrated multi-disciplinary biomarkers discovery and validation study centered at the University of Pittsburgh Cancer Institute (UPCI) focused on GI malignancies, specifically colon and pancreatic cancer. Two major Projects are proposed: (1) Nuclear Matrix Proteins for Early Detection of CRC, and (2) Luminex Multianalyte Profiling (LabMAP(r)) and SELDI-TOF-MS Serum Proteomic Profiling for Early Detection of Pancreatic Cancer. Both projects are firmly based on biomarker discoveries and strong preliminary data developed in our current EDRN/BDL grant and include a team of basic cancer researchers and clinicians supported by faculty with requisite expertise in biostatistics/bioinformatics. Project 1 will further characterize, evaluate, and validate a set of unique nuclear matrix proteins (NMPs) which are expressed in CRC. In preliminary data, the lead NMP biomarker, CC2, is elevated in the serum of CRC patients. A key objective of this Project is to undertake a more extensive Phase 1/2 validation study to assess the performance of this biomarker for serum-based early detection of CRC. Other NMPs in our development pipeline, with a varying spectrum of expression in pre-malignant (adenomas) and malignant tissue, will be further characterized and evaluated as potential biomarkers of early disease and/or risk. Project 2 will extend our preliminary data, developed with EDRN supplemental funds, obtained using two cutting-edge proteomic profiling technologies: 1) a unique panel of Luminex xMAP(r) multiplexed serologic biomarkers comprising multiple pancreatic cancer associated cytokines, chemokines, angiogenic and growth factors and their receptors, as well as circulating antibodies against pancreatic antigens; and 2) SELDI-TOF-MS, that both provide robust discrimination, in serum, of pancreatic cancer patients and case-matched controls. This Project will undertake an extensive Phase 1/2 validation study of these profiling technologies for pancreatic cancer early detection and clinical recurrence of disease. Lastly, as a parallel and focused biomarker discovery aim, we will determine the identity of the key diagnostic peptides/proteins that provide disease classification of the SELDI-TOF-MS profiles. These research Projects will be conducted in the context of an integrated UPCI EDRN/BDL, led by the continuing EDRN PI who brings extensive experience in cancer biomarkers and clinical proteomics research. This team is an outstanding and productive group of clinical and translational cancer researchers focused on cancer biomarkers discovery, evaluation, and validation. These efforts will strongly support the research and clinical goals of the EDRN.